Serological and molecular findings during SARS-CoV-2 infection: the first case study in Finland, January to February 2020

Euro surveillance: European communicable disease bulletin

Haveri, A.; Smura, T.; Kuivanen, S.; Osterlund, P.; Hepojoki, J.; Ikonen, N.; Pitkapaasi, M.; Blomqvist, S.; Ronkko, E.; Kantele, A.; Strandin, T.; Kallio-Kokko, H.; Mannonen, L.; Lappalainen, M.; Broas, M.; Jiang, M.; Siira, L.; Salminen, M.; Puumalainen, T.; Sane, J.; Melin, M.; Vapalahti, O.; Savolainen-Kopra, C.

Abstract

The first case of coronavirus disease (COVID-19) in Finland was confirmed on 29 January 2020. No secondary cases were detected. We describe the clinical picture and laboratory findings 3-23 days since the first symptoms. The SARS-CoV-2/Finland/1/2020 virus strain was isolated, the genome showing a single nucleotide substitution to the reference strain from Wuhan. Neutralising antibody response appeared within 9 days along with specific IgM and IgG response, targeting particularly nucleocapsid and spike proteins.

Keywords

Clinical aspects, diagnosis, treatment; Epidemiology

From SARS to COVID-19: A previously unknown SARS- related coronavirus (SARS-CoV-2) of pandemic potential infecting humans – Call for a One Health approach

One Health
Volume 9, June 2020, 100124

Mohamed E. El Zowalaty, and Josef D. Järhult

Abstract

Human coronaviruses continue to pose a threat to human health. The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in December 2019 which causes coronavirus disease-2019 (COVID-19), an acute respiratory disease marked the third introduction of a highly pathogenic coronavirus into the human population in the twenty-first century. This recent emergence of a previously unknown coronavirus in China leads to huge impacts on humans globally. Covid-19 is a challenge to global public health. Here, we discuss the COVID-19 outbreak in a one health context, highlighting the need for the implementation of one health measures and practices to improve human health and reduce the emergence of pandemic viruses.

Kinetics and pattern of viral excretion in biological specimens of two MERS-CoV cases

Journal of Clinical Virology
Volume 61, Issue 2, October 2014, Pages 275-278

Poissya A .Goffard, E. Parmentier-Decrucq, R. Favory, M. Kauv, E. Kipnis, D. Mathieu, B. Guery, The MERS-CoV Biology Group

Abstract

Background

Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging coronavirus involved in severe acute respiratory distress syndrome (ARDS) and rapid renal failure. Hospital outbreak and nosocomial transmission were reported, however, several issues remain on the viral excretion course.

Objectives

Describe the kinetics and pattern of viral excretion in two infected patients.

Study design

After the initial diagnosis, blood, urine, rectal and respiratory samples were collected regularly, aliquoted and stored at −80 °C. Real-time reverse transcriptase polymerase chain reaction assay targeted the UpE and Orf1a regions of the MERS-CoV genome.

Results

In patient 1, who died of refractory ARDS and renal failure, MERS-CoV RNA was detected in pharyngeal and tracheal swabs, as well blood samples and urine samples until the 30th day. Rectal swabs were negative. Patient 2 also developed multiple-organ failure, but survived, with persisting renal insufficiency (creatinine clearance at 30 mL/min) and persistent interstitial syndrome albeit weaned off mechanical ventilation and no longer requiring oxygen. Tracheal aspirations were positive until the 33rd day, while nasopharyngeal swabs were negative. All other biological samples were negative.

Discussion

Lower respiratory tract excretion of MERS-CoV could be observed for more than one month. The most severely ill patient presented an expression of the virus in blood and urine, consistent with a type-1 interferon mediated immunological response impaired in patient 1, but developed by patient 2. These results suggest that infection control precautions must be adequately evaluated in clinical wards and laboratories exposed to MERS-CoV.

Keywords

Coronavirus, MERS-CoV, RNA detection, Kinetic, Respiratory tract infection

MERS-CoV papain-like protease has deISGylating and deubiquitinating activities

Virology
Volumes 450–451, February 2014, Pages 64-70

Anna M. Mielech, Andy Kilianski, Yahira M. Baez-Santos, Andrew D. Mesecar, Susan C. Baker

Abstract

Coronaviruses encode papain-like proteases (PLpro) that are often multifunctional enzymes with protease activity to process the viral replicase polyprotein and deubiquitinating (DUB)/deISGylating activity, which is hypothesized to modify the innate immune response to infection. Here, we investigate the predicted DUB activity of the PLpro domain of the recently described Middle East Respiratory Syndrome Coronavirus (MERS-CoV). We found that expression of MERS-CoV PLpro reduces the levels of ubiquitinated and ISGylated host cell proteins; consistent with multifunctional PLpro activity. Further, we compared the ability of MERS-CoV PLpro and Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) PLpro to block innate immune signaling of proinflammatory cytokines. We show that expression of SARS-CoV and MERS-CoV PLpros blocks upregulation of cytokines CCL5, IFN-β and CXCL10 in stimulated cells. Overall these results indicate that the PLpro domains of MERS-CoV and SARS-CoV have the potential to modify the innate immune response to viral infection and contribute to viral pathogenesis.

Keywords

MERS-CoV, PLpro, DUB activity, Ubiquitin, deISGylating activity, ISG15

Discovery of N-(benzo[1,2,3]triazol-1-yl)-N-(benzyl)acetamido)phenyl) carboxamides as (SARS-CoV) 3CLpro inhibitors...

Bioorganic & Medicinal Chemistry Letters
Volume 23, Issue 22, 15 November 2013, Pages 6172-6177

Mark Turlington, Aspen Chun, Sakshi Tomar, Aimee Eggler, Valerie Grum-Tokars, Jon Jacobs, J. Scott Daniels, Eric Dawson, Adrian Saldanha, Peter Chase, Yahira M. Baez-Santos, Craig W. Lindsley, Peter Hodder, Andrew D. Mesecar, Shaun R. Stauffer

Abstract

Herein we report the discovery and SAR of a novel series of SARS-CoV 3CLpro inhibitors identified through the NIH Molecular Libraries Probe Production Centers Network (MLPCN). In addition to ML188, ML300 represents the second probe declared for 3CLpro from this collaborative effort. The X-ray structure of SARS-CoV 3CLpro bound with a ML300 analog highlights a unique induced-fit reorganization of the S2–S4 binding pockets leading to the first sub-micromolar noncovalent 3CLpro inhibitors retaining a single amide bond.

Keywords

3CLpro, Severe acute respiratory syndrome, SARS, MERS, Coronavirus

Replication of human coronaviruses SARS-CoV, HCoV-NL63 and HCoV-229E is inhibited by the drug FK506

Virus Research
Volume 165, Issue 1, April 2012, Pages 112-117

Javier Carbajo- Lozoya, Marcel A. Müller, Stephan Kallies, Volker Thiel, Christian Drosten, Albrechtvon Brunn

Abstract

Recent research has shown that Coronavirus (CoV) replication depends on active immunophilin pathways. Here we demonstrate that the drug FK506 (Tacrolimus) inhibited strongly the growth of human coronaviruses SARS-CoV, HCoV-NL63 and HCoV-229E at low, non-cytotoxic concentrations in cell culture. As shown by plaque titration, qPCR, Luciferase- and green fluorescent protein (GFP) reporter gene expression, replication was diminished by several orders of magnitude. Knockdown of the cellular FK506-binding proteins FKBP1A and FKBP1B in CaCo2 cells prevented replication of HCoV-NL63, suggesting the requirement of these members of the immunophilin family for virus growth.

Keywords

SARS-CoV, HCoV-NL63, HCoV-229E, FK506, Tacrolimus, Immunophilins, FKBP1A (FKBP12), FKBP1B (FKBP12.6), Inhibition of viral replication

SARS-CoV 3CLpro inhibitory effects of quinone-methide triterpenes from Tripterygium regelii

Bioorganic & Medicinal Chemistry Letters
Volume 20, Issue 6, 15 March 2010, Pages 1873-1876

Young Bae Ryu, Su-Jin Park, Young Min Kim, Ju-Yeon Lee, Woo Duck Seo, Jong Sun Chang, Ki Hun Park, Mun-Chual Rho, Woo Song Lee

Abstract

Quinone-methide triterpenes, celastrol (1), pristimerin (2), tingenone (3), and iguesterin (4) were isolated from Triterygium regelii and dihydrocelastrol (5) was synthesized by hydrogenation under palladium catalyst. Isolated quinone-methide triterpenes (1–4) and 5 were evaluated for SARS-CoV 3CLpro inhibitory activities and showed potent inhibitory activities with IC50 values of 10.3, 5.5, 9.9, and 2.6 μM, respectively, whereas the corresponding 5 having phenol moiety was observed in low activity (IC50 = 21.7 μM). As a result, quinone-methide moiety in A-ring and more hydrophobic E-ring assist to exhibit potent activity. Also, all quinone-methide triterpenes 1–4 have proven to be competitive by the kinetic analysis.

Keyword

SARS-CoV, 3CLpro, Tripterygium regelii, Quinone-methide, Celastrol, Iguesterin

Synthesis, docking studies, and evaluation of pyrimidines as inhibitors of SARS-CoV 3CL protease

Bioorganic & Medicinal Chemistry Letters
Volume 20, Issue 12, 15 June 2010, Pages 3569-3572

R. Ramajayam, Kian- Pin Tan, Hun- Ge Liu, Po- Huang Liang

Abstract

A series of 2-(benzylthio)-6-oxo-4-phenyl-1,6-dihydropyrimidine as SARS-CoV 3CL protease inhibitors were developed and their potency was evaluated by in vitro protease inhibitory assays. Two candidates had encouraging results for the development of new anti-SARS compounds.

Keywords

SARS-CoV, Pyrimidines

TACE antagonists blocking ACE2 shedding caused by the spike protein of SARS-CoV are candidate antiviral compounds

Antiviral Research
Volume 85, Issue 3, March 2010, Pages 551-555

Shiori Haga, Noriyo Nagata, Tadashi Okamura, Norio Yamamoto, Tetsutaro Sata, Naoki Yamamoto, Takehiko Sasazuki, Yukihito Ishizaka

Abstract

Because outbreaks of severe acute respiratory syndrome coronavirus (SARS-CoV) might reemerge, identifying antiviral compounds is of key importance. Previously, we showed that the cellular factor TNF-α converting enzyme (TACE), activated by the spike protein of SARS-CoV (SARS-S protein), was positively involved in viral entry, implying that TACE is a possible target for developing antiviral compounds. To demonstrate this possibility, we here tested the effects of TACE inhibitors on viral entry. In vitro and in vivo data revealed that the TACE inhibitor TAPI-2 attenuated entry of both pseudotyped virus expressing the SARS-S protein in a lentiviral vector backbone and infectious SARS-CoV. TAPI-2 blocked both the SARS-S protein-induced shedding of angiotensin-converting enzyme 2 (ACE2), a receptor of SARS-CoV, and TNF-α production in lung tissues. Since the downregulation of ACE2 by SARS-S protein was proposed as an etiological event in the severe clinical manifestations, our data suggest that TACE antagonists block SARS-CoV infection and also attenuate its severe clinical outcome.

Keywords

SARS-CoV, TACE, ACE2, Shedding

Antibody-mediated synergy and interference in the neutralization of SARS-CoV at an epitope cluster on the spike protein

Biochemical and Biophysical Research Communications
Volume 390, Issue 3, 18 December 2009, Pages 1056-1060

Lilin Zhong, Lia Haynes, Evi Budo Struble, Azaibi Tamin, Maria Luisa Virata-Theimer, Pei Zhang.

Abstract

Incomplete neutralization of virus, especially when it occurs in the presence of excess neutralizing antibody, represents a biological phenomenon that impacts greatly on antibody-mediated immune prophylaxis of viral infection and on successful vaccine design. To understand the mechanism by which a virus escapes from antibody-mediated neutralization, we have investigated the interactions of non-neutralizing and neutralizing antibodies at an epitope cluster on the spike protein of severe acute respiratory syndrome coronavirus (SARS-CoV). The epitope cluster was mapped at the C-terminus of the spike protein; it consists of structurally intertwined epitopes recognized by two neutralizing monoclonal antibodies (mAbs), 341C and 540C, and a non-neutralizing mAb, 240C. While mAb 341C binds to a mostly linear epitope composed of residues 507PAT509 and V349, mAb 240C binds to an epitope that partially overlaps the former by at least two residues (P507 and A508). The epitope corresponding to mAb 540C is a conformational one, involving residues L504 and N505. In neutralization assays, non-neutralizing 240C disrupted virus neutralization by mAb 341C and/or mAb 540C, whereas a combination of mAbs 341C and 540C blocked virus infectivity synergistically. These findings indicate that the epitope cluster on the spike protein may serve as an evolutionarily conserved platform at which a dynamic interplay between neutralizing and non-neutralizing antibodies occurs, thereby determining the outcome of SARS-CoV infection.

Keywords

SARS-CoV, Monoclonal antibody, Neutralization, Epitope

Identification of a New Region of SARS-CoV S Protein Critical for Viral Entry

Journal of Molecular Biology
Volume 394, Issue 4, 11 December 2009, Pages 600-605

Ying Guo, Jennifer Tisoncik, Susanna McReynolds, Michael Farzan, Bellur S. Prabhakar, Thomas Gallagher, Lijun Rong, Michael Caffrey.

Abstract

Infection by severe acute respiratory syndrome coronavirus (SARS-CoV) is initiated by specific interactions between the SARS-CoV spike (S) protein and its receptor ACE2. In this report, we screened a peptide library representing the SARS-CoV S protein sequence using a human immunodeficiency virus-based pseudotyping system to identify specific regions that affect viral entry. One of the 169 peptides screened, peptide 9626 (S residues 217–234), inhibited SARS-CoV S-mediated entry of the pseudotyped virions in 293T cells expressing a functional SARS-CoV receptor (human angiotensin-converting enzyme 2) in a dose-dependent manner (IC50 ∼ 11 μM). Alanine scanning mutagenesis was performed to assess the roles of individual residues within this region of S, which was previously uncharacterized. The effects included significant reductions in expression (K223A), viral incorporation (L218A, I230A, and N232A), and reduced viral entry (L224A, L226A, I228A, T231A, and F233A). Taken together, these results reveal a new region of the S protein that is crucial for SARS-CoV entry.

Keywords

viral entry, SARS-CoV, envelope, mutagenesis, spike

Investigation of the pharmacophore space of Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) NTPase/helicase by dihydroxychromone derivatives

Bioorganic & Medicinal Chemistry Letters
Volume 19, Issue 16, 15 August 2009, Pages 4538-4541

Chaewoon Lee, Jin Moo Lee, Na- Ra Lee, Dong-Eun Kim, Yong-Joo Jeong, You hoon Chong.

Abstract

Aryl diketoacids have been identified as the first SARS-CoV NTPase/helicase inhibitors with a distinct pharmacophore featuring an arylmethyl group attached to a diketoacid. In order to search for the pharmacophore space around the diketoacid core, three classes of dihydroxychromone derivatives were prepared. Based on SAR study, an extended feature of the pharmacophore model of SARS-CoV NTPase/helicase was proposed which is constituted of a diketoacid core, a hydrophobic arylmethyl substituent, and a free catechol unit.

Keywords

SARS (Severe Acute Respiratory Syndrome), NTPase/helicase, Dihydroxychromone, Pharmacophore

Lipid rafts are involved in SARS-CoV entry into Vero E6 cells

Biochemical and Biophysical Research Communications
Volume 369, Issue 2, 2 May 2008, Pages 344-349

Yanning Lu, Ding Xiang Liu, James P. Tam.

Abstract

Lipid rafts often serve as an entry site for certain viruses. Here, we report that lipid rafts in Vero E6 cells are involved in the entry of severe acute respiratory syndrome coronavirus (SARS-CoV). Infectivity assay showed the integrity of lipid rafts was required for productive infection of pseudotyped SARS-CoV. Depletion of plasma membrane cholesterol with MβCD relocalized raft-resident marker caveolin-1 as well as SARS-CoV receptor ACE2 to a nonraft environment, but did not significantly change the surface expression of ACE2. MβCD-treatment inhibited infectivity of pseudotyped SARS-CoV by 90%. Biochemical fractionation and confocal imaging confirmed that ACE2 colocalized with raft-resident markers. Furthermore, an ectodomain of SARS-CoV S protein (S1188HA) could associate with lipid rafts after binding to its receptor, and colocalize with raft-resident marker ganglioside GM1. The binding of S1188HA was not affected by depleting plasma membrane cholesterol. Taken together, our results support that lipid rafts serve as an entry port for SARS-CoV.

Keywords

Lipid rafts, SARS-CoV, Vero E6, ACE2, Spike protein, Entry

Utility of the aged BALB/c mouse model to demonstrate prevention and control strategies for Severe Acute Respiratory Syndrome coronavirus (SARS-CoV)

Vaccine
Volume 25, Issue 12, 8 March 2007, Pages 2173-2179

Leatrice N. Vogel, Anjeanette Roberts, Christopher D. Paddock, Gillian L. Genrich, Elaine W. Lamirande, Sagar U. Kapadia, John K. Rose, Sherif R. Zaki, Kanta Subbarao.

Abstract

The causative agent of Severe Acute Respiratory Syndrome (SARS) was identified as a coronavirus (CoV) following the outbreak of 2002–2003. There are currently no licensed vaccines or treatments for SARS-CoV infections. Potential prevention and control strategies that show promise in vitro must be evaluated in animal models. The aged BALB/c mouse model for SARS supports a high level of viral replication in association with clinical illness and disease that mimics SARS in the elderly. We tested two preventive strategies, vaccination and passive transfer of serum antibody, to determine the extent of protection achieved against SARS-CoV challenge in this model. These approaches were able to achieve or induce antibody titers sufficient to reduce viral load, protect from weight loss and reduce or eliminate histopathologic changes in the lungs of aged mice. This study validates the utility of the aged BALB/c mouse model for evaluation of the efficacy of vaccines and immunoprophylaxis.

Keywords

SARS-CoV, Aged mouse model, Prophylaxis

Accelerated induction of apoptosis in insect cells by baculovirus-expressed SARS-CoV membrane protein

FEBS Letters
Volume 580, Issue 16, 10 July 2006, Pages 3829-3834

Edited by Hans-Dieter Klenk

Chia-Wei Lai, Zun-Ren Chan, Ding-Gang Yang, Wen-Hsin Lo, Yiu-Kay Lai, Margaret Dah-Tsyr Chang, Yu-Chen Hu

Abstract

It has been shown that severe acute respiratory syndrome-associated coronavirus (SARS-CoV) 3a and 7a proteins, but not membrane (M) protein, induce apoptosis in mammalian cells. Upon expression of SARS-CoV M protein using the baculovirus/insect cell expression system, however, we found that the expressed M protein triggered accelerated apoptosis in insect cells, as characterized by rapid cell death, elevated cytotoxicity, cell shrinkage, nuclear condensation and DNA fragmentation. Conversely, the M protein expressed in mammalian cells did not induce apoptosis. This is the first report describing the induction of apoptosis by SARS-CoV M protein in animal cells and possible implications are discussed.

Keywords

Apoptosis, Baculovirus, Insect cell, Membrane protein, SARS-CoV

Interferon-beta and interferon-gamma synergistically inhibit the replication of severe acute respiratory syndrome-associated coronavirus (SARS-CoV)

Virology
Volume 329, Issue 1, 10 November 2004, Pages 11-17

Bruno Sainz Jr. , Eric C. Mossel, C. J. Peters, Robert F. Garry

Abstract

Recent studies have shown that interferon-gamma (IFN-γ) synergizes with IFN-α/β to inhibit the replication of both RNA and DNA viruses. We investigated the effects of IFNs on the replication of two strains of severe acute respiratory syndrome-associated coronavirus (SARS-CoV). While treatment of Vero E6 cells with 100 U/ml of either IFN-β or IFN-γ marginally reduced viral replication, treatment with both IFN-β and IFN-γ inhibited SARS-CoV plaque formation by 30-fold and replication by 3000-fold at 24 h and by > 1 × 105-fold at 48 and 72 h post-infection. These studies suggest that combination IFN treatment warrants further investigation as a treatment for SARS.

Keywords

Coronavirus, Interferon-gamma, Interferon-beta

Orchitis in a Cat Associated with Coronavirus Infection

Journal of Comparative Pathology
Volume 124, Issues 2–3, February 2001, Pages 219-222

O. G. Sigurdardottir, Ø. Kolbjørnsen and H. Lutz

Abstract

A case of severe, pyogranulomatous and necrotizing orchitis in a cat, which later succumbed to systemic feline infectious peritonitis (FIP), is described. The 3·5-year-old cat, positive for feline immunodeficiency virus infection, presented with a left testicular enlargement. A few months after castration the animal was humanely destroyed due to declining health. Post-mortem examination revealed inflammatory lesions in abdominal organs and in the brain compatible with FIP. Infection was confirmed with a reverse transcriptase–polymerase chain reaction test and by immunohistochemical demonstration of coronavirus antigen in the affected tissues, including the left testicle. FIP is usually a systemic disease. However, lesions and presenting clinical signs in a single organ system such as the brain are not uncommon. The results of this case study indicate that orchitis, although rare, should be on the list of lesions of FIP.

Keywords

coronavirus antigen, orchitis, feline infectious peritonitis,

Release of Coronavirus E Protein in Membrane Vesicles from Virus-Infected Cells and E Protein-Expressing Cells

Virology
Volume 263, Issue 2, 25 October 1999, Pages 265-272

Junko Maeda, Akihiko Maeda, Shinji Makino

Abstract

Coronavirus E protein is a small viral envelope protein that plays an essential role in coronavirus assembly; coexpression of coronavirus M and E proteins results in the production of virus-like particles. The present study demonstrated that mouse hepatitis virus (MHV) E protein was released as an integral membrane protein in lipid vesicles from E-protein-expressing mammalian cells, in the absence of other MHV proteins. Furthermore, our data indicated that the E-protein-containing vesicles, which had a slightly lighter buoyant density than that of MHV, were released from MHV-infected cells. These data implied that E protein alone can drive the production and release of coronavirus envelope in the absence of M protein.

Keywords

Coronavirus, E protein, mouse hepatitis virus